Abstract
The objective of this study was to assess the effects of some culture conditions [temperature (20, 30, 37 °C), incubation time (48, 72, 120 h), pH (5.0, 6.0, 7.0), NaCl concentration (0, 3, 6%), carbon (glucose, fructose, lactose), nitrogen (sodium nitrate, ammonium sulfate, bacto-peptone), and mineral sources (calcium carbonate, ferric chloride)] on the exopolysaccharide (EPS) production by lactic acid bacteria (LAB) strains (belonging to Lactobacillus (L.) plantarum, L. namurensis, and Pediococcus (P.) ethanolidurans species) isolated from naturally fermented pickles. The maximum EPS production was determined at 30 °C and pH 6.0. The highest amount of EPS was obtained after 120 h of incubation, with glucose as carbon source, bacto-peptone as nitrogen source and calcium carbonate as mineral source for most of the tested strains. The EPS formation was not stimulated by NaCl, indicating that EPS formation of the tested strains was not a stress response. L. plantarum MF460 produced the highest amount of EPS at 30 °C after 48 h of incubation, which was 515.48 mg/L. One of the most pronounced results of this study was that the EPS production of L. plantarum MF556 strain was increased up to 512.81 mg/L with the addition of calcium carbonate to MRS medium. The effect of different culture conditions, particularly of incubation time, carbon, nitrogen, and mineral sources, on the EPS production often vary depending on the strain. Therefore, these apparent strain specific results demonstrated that the optimum culture conditions for the enhanced EPS production should be specifically determined for each LAB strain.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abedfar A, Hosseininezhad M, Sadeghi A, Raeisi M, Feizy J (2018) Investigation on "spontaneous fermentation" and the productivity of microbial exopolysaccharides by Lactobacillus plantarum and Pediococcus pentosaceus isolated from wheat bran sourdough. Lwt-Food Sci Technol 96:686–693. https://doi.org/10.1016/j.lwt.2018.05.071
Aslim B, Yuksekdag ZN, Beyatli Y, Mercan N (2005) Exopolysaccharide production by Lactobacillus delbruckii subsp bulgaricus and Streptococcus thermophilus strains under different growth conditions. World J Microb Biot 21:673–677. https://doi.org/10.1007/sl1274-004-3613-2
Aslim B, Beyatli Y, Yuksekdag ZN (2006) Productions and monomer compositions of exopolysaccharides by Lactobacillus delbrueckii subsp bulgaricus and Streptococcus thermophilus strains isolated from traditional home-made yoghurts and raw milk. Int J Food Sci Technol 41:973–979. https://doi.org/10.1111/j.1365-2621.2005.01155.x
Bengoa AA, Llamas MG, Iraporda C, Duenas MT, Abraham AG, Garrote GL (2018) Impact of growth temperature on exopolysaccharide production and probiotic properties of Lactobacillus paracasei strains isolated from kefir grains. Food Microbiol 69:212–218. https://doi.org/10.1016/j.fm.2017.08.012
Caggianiello G, Kleerebezem M, Spano G (2016) Exopolysaccharides produced by lactic acid bacteria: from health-promoting benefits to stress tolerance mechanisms. Appl Microbiol Biot 100:3877–3886. https://doi.org/10.1007/s00253-016-7471-2
Cerning J, Renard CMGC, Thibault JF, Bouillanne C, Landon M, Desmazeaud M, Topisirovic L (1994) Carbon source requirements for exopolysaccharide production by Lactobacillus-Casei Cg11 and partial structure-analysis of the polymer. Appl Environ Microb 60:3914–3919
Cirrincione S, Breuer Y, Mangiapane E, Mazzoli R, Pessione E (2018) 'Ropy' phenotype, exopolysaccharides and metabolism: study on food isolated potential probiotics. LAB Microbiol Res 214:137–145. https://doi.org/10.1016/j.micres.2018.07.004
De Vuyst L, Degeest B (1999) Heteropolysaccharides from lactic acid bacteria. Fems Microbiol Rev 23:153–177. https://doi.org/10.1016/S0168-6445(98)00042-4
De Vuyst L, Vanderveken F, Van de Ven S, Degeest B (1998) Production by and isolation of exopolysaccharides from Streptococcus thermophilus grown in a milk medium and evidence for their growth-associated biosynthesis. J Appl Microbiol 84:1059–1068
De Vuyst L, De Vin F, Vaningelgem F, Degeest B (2001) Recent developments in the biosynthesis and applications of heteropolysaccharides from lactic acid bacteria. Int Dairy J 11:687–707. https://doi.org/10.1016/S0958-6946(01)00114-5
Degeest B, Janssens B, De Vuyst L (2001a) Exopolysaccharide (EPS) biosynthesis by Lactobacillus sakei 0–1: production kinetics, enzyme activities and EPS yields. J Appl Microbiol 91:470–477. https://doi.org/10.1046/j.1365-2672.2001.01404.x
Degeest B, Vaningelgem F, De Vuyst L (2001b) Microbial physiology, fermentation kinetics, and process engineering of heteropolysaccharide production by lactic acid bacteria. Int Dairy J 11:747–757. https://doi.org/10.1016/S0958-6946(01)00118-2
Degeest B, Mozzi F, De Vuyst L (2002) Effect of medium composition and temperature and pH changes on exopolysaccharide yields and stability during Streptococcus thermophilus LY03 fermentations. Int J Food Microbiol 79:161–174. https://doi.org/10.1016/S0168-1605(02)00116-2
Dertli E, Mercan E, Arici M, Yilmaz MT, Sagdic O (2016) Characterisation of lactic acid bacteria from Turkish sourdough and determination of their exopolysaccharide (EPS) production characteristics. Lwt-Food Sci Technol 71:116–124. https://doi.org/10.1016/j.lwt.2016.03.030
Elmaci SB, Tokatli M, Dursun D, Ozcelik F, Sanlibaba P (2015) Phenotypic and genotypic identification of lactic acid bacteria isolated from traditional pickles of the Cubuk region in Turkey. Folia Microbiol 60:241–251. https://doi.org/10.1007/s12223-014-0363-x
Ergene E, Avci A (2018) Effects of cultural conditions on exopolysaccharide production by Bacillus sp ZBP4. Tarim Bilim Derg 24:386–393
Forouhi E, Gunn DJ (1983) Some effects of metal-ions on the estimation of reducing sugars in biological media. Biotechnol Bioeng 25:1905–1911. https://doi.org/10.1002/bit.260250717
Grobben GJ et al (1997) Analysis of the exopolysaccharides produced by Lactobacillus delbrueckii subsp. bulgaricus NCFB 2772 grown in continuous culture on glucose and fructose. Appl Microbiol Biot 48:516–521. https://doi.org/10.1007/s002530051089
Grobben GJ et al (1998) Enhancement of exopolysaccharide production by Lactobacillus delbrueckii subsp. bulgaricus NCFB 2772 with a simplified defined medium. Appl Environ Microb 64:1333–1337
Grobben GJ, Boels IC, Sikkema J, Smith MR, De Bont JAM (2000) Influence of ions on growth and production of exopolysaccharides by Lactobacillus delbrueckii subsp bulgaricus NCFB 2772. J Dairy Res 67:131–135. https://doi.org/10.1017/S002202999900391x
Grosu-Tudor SS, Zamfir M (2013) Functional properties of lactic acid bacteria isolated from Romanian fermented vegetables. Food Biotechnol 27:235–248. https://doi.org/10.1080/08905436.2013.811082
Haj-Mustafa M, Abdi R, Sheikh-Zeinoddin M, Soleimanian-Zad S (2015) Statistical study on fermentation conditions in the optimization of exopolysaccharide production by Lactobacillus rhamnosus 519 in skimmed milk base media. Biocatal Agric Biote 4:521–527. https://doi.org/10.1016/j.bcab.2015.08.013
Harutoshi T (2013) Exopolysaccharides of lactic acid bacteria for food and colon health applications. In: Kongo M (ed) Lactic acid bacteria—R & D for food, health and livestock purposes. IntechOpen, New York, pp. 515–538. https://doi.org/10.5772/50839. https://www.intechopen.com/books/lactic-acid-bacteria-r-d-for-food-health-and-livestock-purposes/exopolysaccharides-of-lactic-acid-bacteria-for-food-and-colon-health-applications
Imran MYM et al (2016) Statistical optimization of exopolysaccharide production by Lactobacillus plantarum NTMI05 and NTMI20. Int J Biol Macromol 93:731–745. https://doi.org/10.1016/j.ijbiomac.2016.09.007
Ismail B, Nampoothiri KM (2010) Production, purification and structural characterization of an exopolysaccharide produced by a probiotic Lactobacillus plantarum MTCC 9510. Arch Microbiol 192:1049–1057. https://doi.org/10.1007/s00203-010-0636-y
Khanh TBTD, Thao DTT (2016) Optimal conditions for exopolysaccharide production by Lactobacillus plantarum T10. J Sci Technol 54:40–47
Kim MJ, Seo HN, Hwang TS, Lee SH, Park DH (2008) Characterization of exopolysaccharide (EPS) produced by Weissella hellenica SKkimchi3 isolated from Kimchi. J Microbiol 46:535–541. https://doi.org/10.1007/s12275-008-0134-y
Li W et al (2014) Production of exopolysaccharides by Lactobacillus helveticus MB2-1 and its functional characteristics in vitro. Lwt-Food Sci Technol 59:732–739. https://doi.org/10.1016/j.lwt.2014.06.063
Liu AP et al (2018) Diversity of isolated lactic acid bacteria in Ya'an sourdoughs and evaluation of their exopolysaccharide production characteristics. Lwt-Food Sci Technol 95:17–22. https://doi.org/10.1016/j.lwt.2018.04.061
Llamas-Arriba MG, Perez-Ramos A, Puertas AI, Lopez P, Duenas MT, Prieto A (2018) Characterization of Pediococcus ethanolidurans CUPV141: a beta-d-glucan- and heteropolysaccharide-producing bacterium. Front Microbiol. https://doi.org/10.3389/fmicb.2018.02041
Looijesteijn PJ, van Casteren WHM, Tuinier R, Doeswijk-Voragen CHL, Hugenholtz J (2000) Influence of different substrate limitations on the yield, composition and molecular mass of exopolysaccharides produced by Lactococcus lactis subsp cremoris in continuous cultures. J Appl Microbiol 89:116–122. https://doi.org/10.1046/j.1365-2672.2000.01082.x
Macedo MG, Lacroix C, Gardner NJ, Champagne CP (2002) Effect of medium supplementation on exopolysaccharide production by Lactobacillus rhamnosus RW-9595M in whey permeate. Int Dairy J 12:419–426. https://doi.org/10.1016/S0958-6946(01)00173-X
Maeda H, Zhu X, Suzuki S, Suzuki K, Kitamura S (2004) Structural characterization and biological activities of an exopolysaccharide kefiran produced by Lactobacillus kefiranofaciens WT-2B. J Agr Food Chem 52:5533–5538. https://doi.org/10.1021/jf049617g
Pan DD, Mei XM (2010) Antioxidant activity of an exopolysaccharide purified from Lactococcus lactis subsp lactis 12. Carbohyd Polym 80:908–914. https://doi.org/10.1016/j.carbpol.2010.01.005
Pham PL, Dupont I, Roy D, Lapointe G, Cerning J (2000) Production of exopolysaccharide by Lactobacillus rhamnosus R and analysis of its enzymatic degradation during prolonged fermentation. Appl Environ Microb 66:2302–2310. https://doi.org/10.1128/Aem.66.6.2302-2310.2000
Polak-Berecka M, Wasko A, Kubik-Komar A (2014) Optimization of culture conditions for exopolysaccharide production by a probiotic strain of Lactobacillus rhamnosus E/N. Pol J Microbiol 63:253–257
Razack SA, Velayutham V, Thangavelu V (2013) Medium optimization for the production of exopolysaccharide by Bacillus subtilis using synthetic sources and agro wastes. Turk J Biol 37:280–288. https://doi.org/10.3906/biy-1206-50
Shin JS et al (2016) Exopolysaccharide fraction from Pediococcus pentosaceus KFT18 induces immunostimulatory activity in macrophages and immunosuppressed mice. J Appl Microbiol 120:1390–1402. https://doi.org/10.1111/jam.13099
Torino MI, Hebert EM, Mozzi F, de Valdez GF (2005) Growth and exopolysaccharide production by Lactobacillus helveticus ATCC 15807 in an adenine-supplemented chemically defined medium. J Appl Microbiol 99:1123–1129. https://doi.org/10.1111/j.1365-2672.2005.02701.x
Torino MI, de Valdez GF, Mozzi F (2015) Biopolymers from lactic acid bacteria. Novel applications in foods and beverages Front Microbiol. https://doi.org/10.3389/fmicb.2015.00834
Tsuda H, Miyamoto T (2010) Production of exopolysaccharide by Lactobacillus plantarum and the prebiotic activity of the exopolysaccharide. Food Sci Technol Res 16:87–92. https://doi.org/10.3136/fstr.16.87
Vandenberg DJC et al (1995) Production of a novel extracellular polysaccharide by Lactobacillus-Sake 0–1 and characterization of the polysaccharide. Appl Environ Microb 61:2840–2844
Vaningelgem F, Zamfir M, Adriany T, De Vuyst L (2004) Fermentation conditions affecting the bacterial growth and exopolysaccharide production by Streptococcus thermophilus ST 111 in milk-based medium. J Appl Microbiol 97:1257–1273. https://doi.org/10.1111/j.1365-2672.2004.02418.x
Velasco S, Arskold E, Paese M, Grage H, Irastorza A, Radstrom P, van Niel EWJ (2006) Environmental factors influencing growth of and exopolysaccharide formation by Pediococcus parvulus 2.6. Int J Food Microbiol 111:252–258. https://doi.org/10.1016/j.ijfoodmicro.2006.06.008
Zhang YC, Li SY, Zhang CH, Luo YK, Zhang HP, Yang ZA (2011) Growth and exopolysaccharide production by Lactobacillus fermentum F6 in skim milk. Afr J Biotechnol 10:2080–2091
Zhang L et al (2013) Antioxidant activity of an exopolysaccharide isolated from Lactobacillus plantarum C88. Int J Biol Macromol 54:270–275. https://doi.org/10.1016/j.ijbiomac.2012.12.037
Zivkovic M, Miljkovic M, Ruas-Madiedo P, Strahinic I, Tolinacki M, Golic N, Kojic M (2015) Exopolysaccharide production and ropy phenotype are determined by two gene clusters in putative probiotic strain Lactobacillus paraplantarum BGCG11. Appl Environ Microb 81:1387–1396. https://doi.org/10.1128/Aem.03028-14
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Communicated by Erko Stackebrandt.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Mıdık, F., Tokatlı, M., Bağder Elmacı, S. et al. Influence of different culture conditions on exopolysaccharide production by indigenous lactic acid bacteria isolated from pickles. Arch Microbiol 202, 875–885 (2020). https://doi.org/10.1007/s00203-019-01799-6
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00203-019-01799-6